Journal:

Article Title: Up-regulation of granulomatous inflammation in interleukin-6 knockout mice infected with Rhodococcus aurantiacus

doi: 10.1111/j.1365-2567.2003.01762.x

Figure Lengend Snippet: Kinetics of endogenous IL-6 production from days 1–28 p.i. in spleen extracts of R. aurantiacus-infected IL-6+/+ mice and IL-6−/− mice. IL-6 was produced during infection with maximal production on day 3 p.i. No IL-6 was detected in spleen extracts of the IL-6−/− mice. Each point represents the mean ± SD for 6–10 mice.

Article Snippet: Administration of recombinant mouse IL-6 (rIL-6) One µg of rIL-6 (R&D Systems, Inc., MN) was administered intravenously to IL-6 −/− mice at day −1of R. aurantiacus infection.

Techniques: Infection, Produced

Journal:

Article Title: Up-regulation of granulomatous inflammation in interleukin-6 knockout mice infected with Rhodococcus aurantiacus

doi: 10.1111/j.1365-2567.2003.01762.x

Figure Lengend Snippet: Histological appearance of livers of IL-6+/+ mice (a), IL-6−/− mice (b), IL-6−/− mice treated with anti-IFN-γ mAb (c), anti-TNF-α mAb (d), and rIL-6 (e) at 2 weeks p.i. A non-necrotic and epithelioid granuloma is seen in the liver of an IL-6+/+ mouse (a), while a large granulomatous lesion with a central area of necrosis is developing in the tissue of IL-6−/− mouse (b). Treatment of IL-6−/− mice with either anti-IFN-γ mAb (c) or anti-TNF-α mAb (d) caused extensive decreases in both number and size of granulomas, and no necrotic granulomas are observed. Administration of rIL-6 reduced the area of necrotic granulomas in the IL-6−/− mouse (e). (original magnification, a, ×400; b–e, ×100).

Article Snippet: Administration of recombinant mouse IL-6 (rIL-6) One µg of rIL-6 (R&D Systems, Inc., MN) was administered intravenously to IL-6 −/− mice at day −1of R. aurantiacus infection.

Techniques:

Journal:

Article Title: Up-regulation of granulomatous inflammation in interleukin-6 knockout mice infected with Rhodococcus aurantiacus

doi: 10.1111/j.1365-2567.2003.01762.x

Figure Lengend Snippet: Granuloma formation in the liver at 2 weeks p.i. The mean area of granulomas per field was determined in 10 optical fields of each section from livers of mice treated in the following ways: IL-6+/+ mice; IL-6−/− mice; IL-6−/− mice treated with anti-IFN-γ mAb; IL-6−/− mice treated with anti-TNF-α mAb; and IL-6−/− mice administered rIL-6.

Article Snippet: Administration of recombinant mouse IL-6 (rIL-6) One µg of rIL-6 (R&D Systems, Inc., MN) was administered intravenously to IL-6 −/− mice at day −1of R. aurantiacus infection.

Techniques:

Journal:

Article Title: Up-regulation of granulomatous inflammation in interleukin-6 knockout mice infected with Rhodococcus aurantiacus

doi: 10.1111/j.1365-2567.2003.01762.x

Figure Lengend Snippet: Kinetics of bacterial number (Log CFU) of R. aurantiacus-infected IL-6+/+ mice, IL-6−/− mice, IL-6−/− mice treated with anti-IFN-γ mAb, and IL-6−/− mice treated with anti-TNF-α mAb. Four groups of mice were infected with 1 × 108 CFU of R. aurantiacus, and the numbers of viable bacteria present in spleens (a) and livers (b) were determined from days 1–14 p.i. Each point represents the mean ± SD of the R. aurantiacus CFU from 10 mice per time point.

Article Snippet: Administration of recombinant mouse IL-6 (rIL-6) One µg of rIL-6 (R&D Systems, Inc., MN) was administered intravenously to IL-6 −/− mice at day −1of R. aurantiacus infection.

Techniques: Infection, Bacteria

Journal:

Article Title: Up-regulation of granulomatous inflammation in interleukin-6 knockout mice infected with Rhodococcus aurantiacus

doi: 10.1111/j.1365-2567.2003.01762.x

Figure Lengend Snippet: Endogenous IFN-γ production in spleen extracts (a) and liver extracts (b) of various mouse groups: infected IL-6+/+ mice, infected IL-6−/− mice, infected IL-6−/− mice treated prophylactically with anti-TNF-α mAb and infected IL-6−/− mice treated prophylactically with rIL-6. Compared with IL-6+/+ mice, a significantly higher level of IFN-γ is observed in the early phase of infection in IL-6−/− mice, whereas IFN-γ production shows a greater decrease at 2 weeks p.i. in IL-6−/− mice. Treatment with anti-TNF-α mAb prevented IFN-γ production. In the IL-6−/− mice treated with rIL-6, the production of IFN-γ in the early phase is observed to decrease. The data are the mean ± SD of 6 mice per group *P < 0·01.

Article Snippet: Administration of recombinant mouse IL-6 (rIL-6) One µg of rIL-6 (R&D Systems, Inc., MN) was administered intravenously to IL-6 −/− mice at day −1of R. aurantiacus infection.

Techniques: Infection

Journal:

Article Title: Up-regulation of granulomatous inflammation in interleukin-6 knockout mice infected with Rhodococcus aurantiacus

doi: 10.1111/j.1365-2567.2003.01762.x

Figure Lengend Snippet: Endogenous TNF-α production in liver extracts of various mouse groups: infected IL-6+/+ mice, infected IL-6−/− mice, infected IL-6−/− mice treated prophylactically with anti-IFN-γ mAb and infected IL-6−/− mice treated prophylactically with rIL-6. Compared with IL-6+/+ mice, a significantly higher level of TNF-α is observed in the early phase of infection in IL-6−/− mice. Treatment with either anti-IFN-γ mAb or rIL-6 prevented TNF-α production. The data are the mean ± SD of 6 mice per group *P < 0·01.

Article Snippet: Administration of recombinant mouse IL-6 (rIL-6) One µg of rIL-6 (R&D Systems, Inc., MN) was administered intravenously to IL-6 −/− mice at day −1of R. aurantiacus infection.

Techniques: Infection

Journal: Clinical science (London, England : 1979)

Article Title: DAPT, a potent Notch inhibitor regresses actively growing abdominal aortic aneurysm via divergent pathways

doi: 10.1042/CS20200456

Figure Lengend Snippet: (A–F) Expression of pro-inflammatory genes (Il6, Il12, and iNos; A–C) and anti-inflammatory genes (Arg1, c-Myc, and Egr2; D–F) with vehicle or DAPT (10 μM) treatment at 6 h (A–C) and 12 h (D–F) of low LPS (10 ng/ml + IFN-γ 10 ng/ml) stimulation in Apoe−/− BMDMs. (G) Measurement of soluble IL6 in the supernatant of macrophages treated with low LPS for 12 h in presence of DAPT (10 μM) as measured by ELISA. (H–J) Gene expression of Adam17 (H), gp130 (I), and Stat3 (J) in macrophages treated with low LPS. Briefly, cells were treated with LPS for 3 h, washed and then further treated with DAPT for 6 h for gene expression analysis. (K) Representative images showing phagocytosis of zymosan particles by BMDMs in various conditions. (L) Quantitative analysis of the phagocytosis. (M) NICD and IL6 expression and their co-localization in the aorta of healthy and AAA human patients as determined by immunofluorescence. (N) Quantification of relative fluorescence intensity of IL6 and NICD. *P<0.05, **P<0.01, ***P<0.001 in paired two-tailed Student’s t test.

Article Snippet: BMDMs were serum-starved and then treated with either low LPS or vehicle for 3 h. Thereafter, cells were washed and treated with either DAPT, IL6 antibody (1:1000; ab6672) or recombinant IL6 protein (25 ng/ml, R&D systems) for 24 h. Cells were washed and incubated with fluorescein-conjugated Zymosan A bioparticles (20 particles/cell, Invitrogen) for 1 h. Unengulfed bioparticles were removed by washing with cold PBS.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Fluorescence, Two Tailed Test

Journal: Clinical science (London, England : 1979)

Article Title: DAPT, a potent Notch inhibitor regresses actively growing abdominal aortic aneurysm via divergent pathways

doi: 10.1042/CS20200456

Figure Lengend Snippet: (A–D) mRNA expression of pro-inflammatory cytokines (Il6, Il12, iNos, and Cd38) in the aorta of experimental mice (n=3). (E–H) mRNA expression of anti-inflammatory cytokines (Arg1, Mgl2, cMyc, and Egr2) in the aorta of experimental mice (n=3). (I,J) Representative IHC images showing immunoreactivity of F4/80 and Il6 in the suprarenal aorta. (K,L) Quantification of F4/80 and Il6 immunostaining (n=6). Paired two-tailed Student’s t test was performed for (A–H) and ANOVA followed by Tukey’s multiple comparison analysis was performed for (K,L). *P<0.05; **P<0.01; ***P<0.001; Scale bar = 50 μm. Abbreviation: lm, lumen.

Article Snippet: BMDMs were serum-starved and then treated with either low LPS or vehicle for 3 h. Thereafter, cells were washed and treated with either DAPT, IL6 antibody (1:1000; ab6672) or recombinant IL6 protein (25 ng/ml, R&D systems) for 24 h. Cells were washed and incubated with fluorescein-conjugated Zymosan A bioparticles (20 particles/cell, Invitrogen) for 1 h. Unengulfed bioparticles were removed by washing with cold PBS.

Techniques: Expressing, Immunostaining, Two Tailed Test, Comparison

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: Effect of exogenous IL6 on caloric intake and body mass. Wild-type mice were fed on chow or high-fat diet (HFD) and treated intraperitoneally (IP) with exogenous IL6 or saline as depicted in A. Body mass ( B , D ) and caloric intake ( C , E ) were determined in mice fed chow ( B , C ) and HFD ( D , E ). In all experiments, n = 5 mice/group. Data was analyzed using two-way ANOVA with repeated measures and Bonferroni post-test. ** p < 0.01; *** p < 0.001

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: Hypothalamic transcript expression of IL6. In order to identify obese-prone ( H ) and obese-resistant ( L ) mice, we performed the procedure as depicted in A. The expression of hypothalamic IL6 was determined using real-time PCR ( B ). Palmitate was injected in a single intracerebroventricular (ICV) dose and hypothalamus was collected ( C ) for determinations of IL6 ( D ), TNFα ( E ), and IL1β ( F ) transcripts. B , n = 7–8 mice/group; D – F , n = 5 mice/group; * p < 0.05, ** p < 0.01, *** p < 0.001. In bar graphs, results are presented as mean ± standard deviation

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Injection, Standard Deviation

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: IL6 effect on hypothalamic cell proliferation and survival. Mice were treated with IL6 and BrdU according to the protocols as depicted in A and D . The mediobasal hypothalamus was prepared for analysis using immunofluorescence and confocal microscopy; cell counting was performed using 20× and 40× magnifications. Representative images are presented in B (proliferation assay) and E (survival assay). Total BrdU countings are presented in C (proliferation assay) and F (survival assay). In both assays, n = 6 mice/group. In bar graphs, results are presented as mean ± standard deviation

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: Immunofluorescence, Confocal Microscopy, Cell Counting, Proliferation Assay, Clonogenic Cell Survival Assay, Standard Deviation

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: IL6 induces neurogenesis in the hypothalamus. Mice were treated with a single intraperitoneal dose of IL6, according to the protocol as depicted in A . The hypothalamic expressions of Sox6 ( B ) and Sox2 ( C ) were determined using real-time PCR. The transcript expressions of Sox6 ( D ) and doublecortin ( E ) were determined in the hypothalami collected from obese-prone ( H ) and obese-resistant ( L ) mice and treated with a single dose of IL6. F Mice were treated with IL6 and BrdU according to the protocols as depicted in Fig. . The mediobasal hypothalamus was prepared for analysis using immunofluorescence and confocal microscopy; cell counting was performed using 20× and 40× magnifications. Total BrdU/NeuN counting is depicted in G (neurogenesis assay). B , C n = 5 mice/group; D , E n = 8–10 mice/group; F , G , n = 5 mice/group. * p < 0.05, ** p < 0.01. In bar graphs, results are presented as mean ± standard deviation

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: Real-time Polymerase Chain Reaction, Immunofluorescence, Confocal Microscopy, Cell Counting, Standard Deviation

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: Analysis of single-cell RNA sequencing data reveal the expression pattern of IL6-receptor encoding genes in the arcuate nucleus of the hypothalamus. A Diffusion-based manifold approximation and projection (dbMAP) embedding of 20,921 single-cell transcriptomes of the arcuate nucleus of the hypothalamus, colored by their annotated cluster of origin. In this embedding, similar cells are mapped tightly, whereas dissimilar cells are mapped apart from each other, rendering clusters of cell types. Cells were annotated by their main cell type as defined in the original study. B (top) Il6ra mRNA expression, key-colored by expression intensity; Il6ra is found mainly in perivascular macrophages/microglia and ependymocytes and also in astrocytes and endothelial cells. B (bottom) Cx3cr1 (a canonical microglial marker) mRNA expression, key-colored by expression intensity. C (top) Il6st mRNA expression, key-colored by expression intensity; Il6st is broadly expressed in the arcuate with some overlapping with the expression pattern of Ntrk2 (bottom), which encodes a neurotrophic receptor. D Violin plot of the mRNA expressions of Il6, Il6ra, Il6st, Cx3cr1, Vim, Ntrk2, and Gfap in the arcuate hypothalamus, per main cell types. Oligodend, oligodendrocyte; NG2/OPC, oligodendrocyte progenitor cells; PVMMicro, perivascular macrophages/microglia; VLMCs, vascular and leptomeningeal cells; ParsTuber, pars tuberalis

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: RNA Sequencing Assay, Expressing, Diffusion-based Assay, Marker

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: IL6 deficiency predisposes to obesity. IL6 was determined in the serum of wild-type (WT) and IL6 knockout (KO) mice treated or not with 1.0 ng exogenous IL6 ( A ). Body mass was determined at age 8 weeks ( B ). The expression of transcripts encoding for hypothalamic neurotransmitters ( C , D ) and neurogenesis-related genes ( E , F ) were determined using real-time PCR in hypothalamic samples collected in mice aged 7 days ( C , E ) or 8 weeks ( D , F ). A n = 3–4 mice/group; n = 25 mice/group; C , E n = 5–7 mice/group; D , F n = 9 mice/group. * p < 0.05, *** p < 0.001. In bar graphs, results are presented as mean ± standard deviation

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: Knock-Out, Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: IL6 induce neurogenic factors expression in hypothalamic neuroprogenitor cells (NPCs). Transcripts were determined using real-time PCR in the NPCs of WT and IL6 KO mice after a 5-day differentiation period: first, WT and IL6 KO hypothalamic cells were analyzed in basal conditions ( A ), and then, IL6 KO NPCs were treated with exogenous IL6 in doses depicted ( B ). Representative immunocytochemistry images of 7-day differentiated cells (green, DCX; red, GFAP; blue, DAPI) ( C ) and DCX-positive cell counting ( D ), and representative immunocytochemistry images of 14-day differentiated cells (green, MAP2; red, GFAP; blue, DAPI) ( E ) and MAP2-positive cell counting ( F ). After 10- ( G , I , K – M ) or 14-day ( H and J ) differentiation period, transcripts were determined using real-time PCR in the NPCs of WT and IL6 KO mice treated with exogenous IL6 in doses depicted in the panels (G-M). A , B n = 12 (3–4 well/group, 2 independent experiments); C – F n = 24 (4 well/group, 2 independent experiments); G – M n = 48 (6–9 well/group, 3 independent experiments). * p < 0.05, ** p < 0.01. In bar graphs, results are presented as mean ± standard deviation

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Immunocytochemistry, Cell Counting, Standard Deviation